The objective of this project is to develop a sensitive and specific assay for prostaglandins in human cerebrospinal fluid (CSF) from patients with alcoholism and appropriate controls using gas chromatography / mass spectrometry. This study was initiated because of literature reports which suggested that a relationship exists between central nervous system effects of ethanol and the central production of prostaglandins. In particular, in human and animal studies, administration of prostaglandin synthesis inhibitors prior to administration of ethanol attenuated central nervous system effects of ethanol. Samples assayed after methodological validation was completed, contained, none of the measured prostaglandins (PGE2, PGE1, PGF2a, PGF1a, 6-keto-PGF1a) at a concentration more than twice the limit of quantification (3 pg/mL CSF). Comparison of GC/MS and radioimmunoassay methods provided further validation for these results. Literature reports of much higher levels of prostaglandins in normal controls, i.e., tens to hundreds of pg/mL CSF, appear to be incorrect. Examination of monkey CSF provided a positive control, since several prostaglandins were easily quantifiable in these samples.